A whole-transcriptome approach to evaluate reference genes for quantitative diurnal gene expression studies under natural field conditions in Tamarix ramosissima leaves

Background: Tamarix ramosissima is a desert forest tree species that is widely distributed in the drought-stricken areas to sustain the fragile ecosystem. Owing to its wide usage in the desert restoration of Asia, it can be used as an ecophysiological model plant. To obtain reliable and accurate results, a set of reference genes should be screened before gene expression. However, up to date, systematical evaluation of reference genes has not been conducted in T. ramosissima. Results: In this study, we used eigenvalues derived from principal component analysis to identify stable expressed genes from 72,035 unigenes from diurnal transcriptomes under natural field conditions. With combined criteria of read counts above 900 and CV of FPKM below 0.3, a total of 7385 unigenes could be qualified as candidate reference genes in T. ramosissima. By using three statistical algorithm packages, geNorm, NormFinder, and BestKeeper, the stabilities of these novel reference genes were further compared with a panel of traditional reference genes. The expression patterns of three aquaporins (AQPs) suggested that at least UBQ (high expression), EIF4A2 (low expression), and GAPDH (moderate expression) could be qualified as ideal reference genes in both RT-PCR and RNA-seq analysis of T. ramosissima. Conclusions: This work will not only facilitate future studies on gene expression and functional analysis of genetic resources of desert plants but also improve our understanding of the molecular regulation of water transport in this plant, which could provide a new clue to further investigate the drought adaptation mechanism of desert plant species under harsh environments.

Phenolic compounds in branches of Tamarix rasissima / 中国中药杂志

To study the chemical constituents of the branches of Tamarix rasissima, repeated silica gel column chromatography, Sephadex LH-20 chromatography and recrystallization were applied for chemical constituents isolation and purification. Ten phenolic compounds were isolated from the n-BuOH fraction and their structures were elucidated by physical properties and spectra analysis such as UV, ESI-MS and NMR as monodecarboxyellagic acid (1), ellagic acid (2), 3, 3'-di-O-methylellagic acid (3), 3, 3'-di-O-methylellagic acid-4-O-beta-D-glucopyranoside (4), 3, 3'-di-O-methylellagic acid-4'-O-alpha-D-arabinfuranoside (5), ferulic acid (6), isoferulic acid (7), caffeic acid (8), 4-O-acetyl-caffeic acid (9), and 4-methyl-1, 2-benzenediol (10). All compounds except for isoferulic acid were isolated firstly from this plant except for isoferulic acid, and compounds 5, 9 and 10 were obtained from Tamarix genus for the first time.

Screening of some ethnobotanically important plants of Nagar area Gilgit-Baltistan, Pakistan

Certain species of ethnobotanical importance belonging to families Asteraceae, Berberidaceae, Cupressaceae, Elaeagnaceae, Gentianaceae, Salicaceae and Tamaricaceae, were collected from Gilgit during the months of June and July 2008 and were investigated for the presence of alkaloids, amino acids, anthraquinone [free and as glycosides] ascorbic acid, carbohydrates, coumarins, flavonoids, phenolics, proteins, saponins and steroids in their aqueous, ethanol and benzene extracts. Artemisia maritime L. [leaves] showed positive results for carbohydrates, coumarins, phenolics and proteins. Ephedra gerardiana Wall ex. Stapf. [stem] tested positive for alkaloids, ascorbic acid, coumarins, phenolics, proteins, saponins and steroids. Tamarix gallica L. tested positive for alkaloids, amino acids, anthraquinone as glycoside, ascorbic acid, carbohydrates, flavonoids, phenolics, proteins and steroids in stems, roots and leaves. Salix acmophylla Boiss. showed positive results for alkaloids, amino acids, anthraquinone [free and as glycosides] ascorbic acid, carbohydrates, flavonoids, phenolics, proteins, saponins and natural steroids. Hippophae rhamnoides L. showed positive results for alkaloids, amino acids, anthraquinone [free as glycosides] ascorbic acid, carbohydrates, coumarins, flavonoids, phenolics, proteins, saponins and steroids. Berberis glycocarpa Stapf. showed positive results for alkaloids, amino acids, ascorbic acid, carbohydrates, flavonoids, phenolics, proteins and steroids in stem, roots and leaves. Similarly Juniperus excelsa Wall ex. C.A. Meyer showed positive result for anthraquinone [both free and as glycosides], carbohydrates, phenolics, proteins, saponins and natural steroids

Study on chemical constituents from ethyl acetate extract of Myricaria bracteata / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical constituents from the ethyl acetate extract of Myricaria bracteata.</p><p><b>METHOD</b>The chemical constituents were isolated and purified by chromatographic techniques, and their structures were identified by physical characters and spectroscopic analysis.</p><p><b>RESULT</b>Sixteen compounds were isolated from the ethyl acetate portion of the 95% ethanolic extract of Myricaria bracteata, and identified as myricarin (1), myricarin B (2), 3alpha-hydroxytaraxer-14-en-28-oic acid (3), myricadiol (4), trans-ferulic acid 22-hydroxydocosanoic acid ester (5), docosyl-3, 4-dihydroxy-trans-cinnamate (6), dillenetin (7), 3, 5, 4'-trihydroxy-7-methoxyflavone (8), 3, 5, 4'-trihydroxy-7, 3'-dimethoxyflavone (9), methyl 3, 5-dihydroxy-4-methoxybenzoate (10), 3-hydroxy-4-methoxy cinnamic acid (11), sinapaldehyde (12), vanillin (13), syringaldehyde (14), 3, 3', 4'-trimethoxyellagic acid (15), methyl p-hyroxybenzoate (16).</p><p><b>CONCLUSION</b>Compounds 5, 6, 12-16 were isolated from the genus Myricaria for the fist time, all of the compounds were isolated from this plant for the fist time, except for 8 and 9.</p>

Chemical constituents from Myricaria alopecuroides / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the chemical constituents in the leaves and branches of Myricaria alopecuroides.</p><p><b>METHOD</b>Solvent extraction method was employed to extract and partition. The chemical constituents were isolated by column chromatography on silica gel, Sephadex LH-20, highly porous resin HP-20. The structures of the compounds were elucidated on the basis of physiochemical properties and spectral analysis.</p><p><b>RESULT</b>Eleven compounds were isolated from this plant and identified as ellagic acid 3,3',4-trimethylether (1), ellagic acid 3,3'-dimethylether (2), isorhamnetin (3), kaempferol (4), 3, 5-dihydroxy-4-methoxybenzoic acid (5), daucosterol (6), 6,7,10-trihydroxy-8-octadecenoic acid (7), quercetin (8), gallic acid (9), palmitic acid (10), hexadecanoic acid, 2,3-dihydroxypropyl ester (11).</p><p><b>CONCLUSION</b>Except 8 and 9, all compounds were isolated from M. alopecuroides for the first time. Compound 1, 2, 5, 7, 10, 11 were obtained from the genus Myricaria for the frist time.</p>

Analysis on Cistanche tubulosa that parasites on different Tamarixs by RAPD / 中国中药杂志

<p><b>OBJECTIVE</b>To study genetic difference of Cistanche tubulosa that parasites on different Tamarixs and give a reference to select host of C. tubulosa.</p><p><b>METHOD</b>Sixteen selected primers by random amplified polymorphic DNA (RAPD) markers were used to analyze genetic distance of C. tubulosa that parasites on eight different hosts.</p><p><b>RESULT</b>Sixty-six point seven percent of the total bands were polymorphic, that proved the genetic diversity level in different C. tubulosa types was relatively high, especially the two that parasites on Tamarix hispida and T. chinensis. Cultural areas had more remarkable influence on genetic distance of Cistanche tubulosa than the hosts, and introduction was helpful to maintain the more genetic diversity in different C. tubulosa types. Genetic difference in different C. tubulosa types was far less than that between different species in Cistanche.</p><p><b>CONCLUSION</b>C. tubulosa types which parasite on different Tamarixs have high genetic diversity.</p>

Study on chemical constituents of Myricaria paniculata I / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical constituents of Myricaria paniculata.</p><p><b>METHOD</b>Silica gel column chromatography was used to separate and purify the chemical constituents, and the structures were elucidated by spectral analysis.</p><p><b>RESULT</b>Four compounds were isolated from the petroleum ether soluble portion, identified as 28-aldehyde-taraxerenone (1), 28-hydroxy-taraxerenone (2), epi-friedelanol (3), 4-methyl stigmast-7-en-3-ol (4). Three compounds were isolated from the EtOAc soluble portion, identified as morelloflavone (5), methyl 3, 5-dihydroxy-4-methoxybenzoate (6), 3-hydroxy-4-methoxy cinnamic acid (7).</p><p><b>CONCLUSION</b>All of these compounds were isolated from the genus for the first time.</p>

Study on secondary metabolic organ of echinacoside in herbs of Cistanche tubulosa / 中国中药杂志

<p><b>OBJECTIVE</b>It could give some theory support of confirming the secondary metabolism organ and regulation of echinacoside in Cistanche tubulosa by searching parasitic growth of C. tubulosa ahd echinacoside variation in different organs of host and parasite.</p><p><b>METHOD</b>The echinacoside content was analyzed by HPLC. The relationship between dry matter accumulation and echinacoside accumulation of C. tubulosa as the well as root diameter of host were comparatively analyzed.</p><p><b>RESULT</b>With the increase of dry matter accumulation of C. tubulosa, echinacoside accumulation increased significantly, and both of them were in significantly positive correlated with the root diameter of host. Echinacoside content in haustorium phloem was 15.53%, higher than that of haustorium xylem, C. tubulosa plant and other organs.</p><p><b>CONCLUSION</b>Haustorium phloem was probably the secondary metabolism organ of echinacoside in C. tubulosa.</p>

Inoculation experiments of Cistanche tubulosa on 8 introduced Tamarix species / 中国中药杂志

<p><b>OBJECTIVE</b>To analyze the inoculation ratio and echinacoside content of Cistanche tubulosa and provide theoretical basis for Tamarix introduction, resource protection and screening of C. tubulosa.</p><p><b>METHOD</b>8 Tamarix species were introduced in the North China Plain and inoculation of C. tubulosa was conducted on all species. Phenylethanoid glycosides fingerprinting and echinacoside content of C. tubulosa were analyzed by using HPLC.</p><p><b>RESULT</b>The adaptability of 8 Tamarix species were significantly different, phenylethanoid glycosides component of C. tubulosa on T. gansuensis and T. austromongolica were basically identical in contrast to T. chinensis, echinacoside content showed no obvious difference in C. tubulosa plant growing 4 months.</p><p><b>CONCLUSION</b>T. gansuensis and T. Austromongolica are suitable for the host introduction plant of C. tubulosa resource protection and screening in North China Plain.</p>

Studies on chemical constituents in herb of Myricaria bracteata / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical constituents of Myricaria bracteata.</p><p><b>METHOD</b>The chemical constituents were isolated by silica gel column chromatography and the structures were elucidated by spectroscopic methods.</p><p><b>RESULT</b>Eleven compounds were obtained and identified as rhamnetin, 3,5,4'-trihydroxy-7,3'-dimethoxyflavone, 3,5,4'-trihydroxy-7-methoxyflavone, quercetin-3-O-alpha-L-rhamnopyranoside, kaempferol, quercetin, chrysoerio, gallic acid, gallic acid ethylester, beta-sitosterol, daucosterol.</p><p><b>CONCLUSION</b>All compounds were obtained from M. bracteata for the first time.</p>

Study on expression of genes in Tamarix androssowii under NaHCO3 stress using gene chip technology / 生物工程学报

Gene chip technology was employed to study gene expression in Tamarix androssowii under NaHCO3 stress. cDNAs from T. androssowii treated with NaHCO3 solution and that from control group were labeled with fluorescent dye CyS and Cy3 respectively. The two fluorescent cDNA probes were mixed and hybridized to gene chips containing T. androssowii genes, and the chips were scanned using biochip scanning system. Differential expression of genes was analyzed through calculation of the ratio of Cy5 to Cy3 signal intensities. Total of 89 genes differentially expressed were identified, among them, 27 showed down regulated expression and 62 showed up regulated expression. Blastx analysis showed that the function of the differentially expressed genes could be grouped into some categorizations such as photosynthesis, reactive oxygen species eliminated, regulation of osmotic potential, regulation of gene expression and signal transduction, metabolism, development, ribosomal protein, protein breakdown and recycling, transporter, water channel proteins and so on. Based on this research, some function-unknown or novel unreported genes that respond to salt stress were also identified, and these genes may have important functions in salt resistance of T. androssowii. Some important pathways of salt resistance in T. androssowii are revealed, and the gene expression profiling of T. androssowii under salt stress and without stress is obtained in this study.

Study on inoculation technology of Cistanche tubulosa in the field / 中国中药杂志

<p><b>OBJECTIVE</b>To increase inoculation rate of Cistanche tubulosa in the field by studying inoculation technologies.</p><p><b>METHOD</b>Root-tube inoculation methed was used on field experiments. Inoculation rate of C. tubulosa was compared to different size seeds and inoculation mediums and inoculation time.</p><p><b>RESULT AND CONCLUSION</b>May is suitable inoculation time. The inoculation rate of C. tubulosa is 92.5% while the seed width is more than 0.7 mm and coarse sand is selected during inoculation period.</p>

Morphology of seed germination and parasitism in Cistanche tubulosa / 中国中药杂志

<p><b>OBJECTIVE</b>To understand the process of Cistanche tubulosa.</p><p><b>METHOD</b>The process of seed germination and parasitism was observed using stereomicroscope.</p><p><b>RESULT</b>Seedling of C. tubulosa sprouted after forty day without host root's contact in fields, a tube-like-organ formed and grew auger-type from host root, the tuber apex where touches host root swelled and formed haustorium. Haustorium intruded host root epidermis and vascular bundles, and released brown substances. Then, embryo bud with six or more young leaves formed, finally the swelled tuber-like-organ broken and seed coat shed. Due to the parasitism of C. tubulosa, the host root near stem site swelled, but the other part, shrunk and disappered gradually.</p><p><b>CONCLUSION</b>Seed of C. tubulosa could germinate indepently in fields. Tuber-like-organ formatin, haustorium formation and bud formation are key steps of C. tubulosa seedling development.</p>

The determination of echinacoside and acteoside in herbs of Cistanche tubulosa / 中国中药杂志

<p><b>OBJECTIVE</b>To analyse the echinacoside and acteoside content of Cistanche tubulosa, collected from different hosts and different size of the cultivated, which is compared to the wilding by RP-HPLC method.</p><p><b>METHOD</b>An Agilent Eclipse XDB-C18(4.6 mm x 250 mm, 5 microm) column was used and a mixture of methanol-acetonitrile-1% acetic acid (15:10:75) was used as the mobile phase at a flow rate of 0.6 mL x min(-1). The column temperature was 30 degrees C and the UV detection wavelength was 334 nm.</p><p><b>RESULT</b>The calibration curves of echinacoside and acteoside were in good linearity over the range of 0.904-9.04 microg (r = 0.999 9), and 1.27-12.7 microg (r = 0.999 9) respectively and the average recoveries of echinacoside (and acteoside) were 98.9% (n = 5, RSD 1.9%), and 97.0% (n = 5, RSD 0.97%).</p><p><b>CONCLUSION</b>The method is simple, quick, acurate. In all of the samples, the contents of echinacoside is markedly more than that of acteoside, the content of the two active component in the wilding is higher than that in the planting. The content of sample in the different sizes gradually increase from the big to the small, and the contents of samples collected from the different hosts vary markedly. These results are useful for the quality evaluation of medicinal materials of C. tubulosa.</p>